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PubMed1994

The Cladophora complex (Chlorophyta): new views based on 18S rRNA gene sequences.

Evolutionary relationships among species traditionally ascribed to the Siphonocladales/Cladophorales have remained unclear due to a lack of phylogenetically informative characters and extensive morphological plasticity resulting in morphological convergence. This study explores some of the diversity within the generic complex Cladophora and its siphonocladalaen allies. Twelve species of Cladophora representing 6 of the 11 morphological sections recognized by van den Hoek were analyzed along with 8 siphonocladalaen species using 18S rRNA gene sequences. The final alignment consisted of 1460 positions containing 92 phylogenetically informative substitutions. Weighting schemes (EOR weighting, combinatorial weighting) were applied in maximum parsimony analysis to correct for substitution bias. Stem characters were weighted 0.66 relative to single-stranded characters to correct for secondary structural constraints. Both weighting approaches resulted in greater phylogenetic resolution. Results confirm that there is no basis for the independent recognition of the Cladophorales and Siphonocladales. The Siphonocladales is polyphyletic, and Cladophora is paraphyletic. All analyses support two principal lineages, of which one contains predominantly tropical members including almost all siphonocladalean taxa, while the other lineage consists of mostly warm- to cold-temperate species of Cladophora.

Base Sequence↗
PubMed1989

The linear 20 kb mitochondrial genome of Pandorina morum (Volvocaceae, Chlorophyta).

A physical restriction map of the mitochondrial genome from one clone (TCC 854) of the sexually isolated populations (syngens) of the morphologically uniform species Pandorina morum Bory has been constructed using restriction endonucleases Ava I, Bam HI, Bgl II, Eco RI, Kpn I, and Pst I. The 20 kb linear genome can easily be separated from plastid DNA, nuclear satellite rDNA, and main band (nuclear) DNA on a Hoechst/CsCl buoyant density gradient. The Pandorina mitochondrial DNA shows sufficient similarity to the 16 kb mitochondrial genome of Chlamydomonas reinhardtii to cross-hybridize, and also hybridizes with a probe containing maize mitochondrial 18S rRNA genes. Double digests, self-probing, and Bal31 exonuclease experiments suggest that 1.8 to 3.3 kb of sequence is repeated at each end of the genome as an inverted repeat. Mitochondrial genome sizes of other P. morum syngens were found to range from ca. 20 to ca. 38 kb. The mitochondrial genome should be valuable for taxonomic studies; it can be used for comparative organellar studies; and it should be of interest to compare with that of other plant and animal mitochondrial genomes.

Biological Evolution↗
PubMed1995

Evolution of nuclear rDNA ITS sequences in the Cladophora albida/sericea clade (Chlorophyta).

Ribosomal DNA ITS sequences were compared among 13 different species and biogeographic isolates from the monophyletic "albida/sericea clade" in the green algal genus Cladophora. Six distinct ITS sequence types were found, characterized by multiple insertions and deletions and high levels of nucleotide substitution. Conserved domains within the ITS regions indicate the presence of ITS secondary structure. Low transition/transversion ratios among the six types and nearly symmetrical tree-length frequency distributions indicate some saturation, and low phylogenetic signal. Although branching order among five of the six ITS sequence types could not be resolved, estimates of ITS sequence divergence as compared with 18S divergence in a subset of the taxa suggests that the origin of the different ITS types is probably in the mid-Miocene (12 Ma ago) but that biogeographic isolates within a single ITS type (including both Pacific and Atlantic representatives) have probably dispersed on a time scale of thousands rather than millions of years.

Base Sequence↗
PubMed1980

The comparative aspects of cell wall chemistry in the green algae (Chlorophyta).

The origin of a cell wall was an event of fundamental importance in the evolution of plants. In the green algae, cell walls apparently had independent origins in at least three lines of evolution. In this paper, the components of the cell wall were determined and compared in four filamentous green algae representing the charophycean, chlorophycean and ulvacean evoluationary lines. The walls of all four have hydroxyproline-containing proteins which separate into five or six bands upon SDS gel electrophoresis. Variation does exist, with the charophyte possessing fast moving electrophoretic bands and high hydroxyproline content, the chlorophytes having intermediate movement of bands and lower hydroxyproline content, and the ulvaecean representative possessing slow moving bands and a very low, if not questionable, hydroxyproline and saccharide content. Qualitative and quantitative estimates of wall proteins and sugars have been determined and compared. A hypothetical scheme of cell wall evolution based on these data, those of previous analyses, and recent phylogenetic schemes is presented. Although sound conclusions cannot be made until more information is available, the scheme might help to emphasize the areas most in need of additional research.

Amino Acids↗
PubMed2003

Effects of copper on wild and tolerant strains of the lichen photobiont Trebouxia erici (Chlorophyta) and possible tolerance mechanisms.

The influence of copper was assessed on wild and tolerant strains of the lichen photobiont Trebouxia erici and shown to have multiple toxic effects. Addition of 4 mM copper chloride into Trebouxia media reduced growth rates of the wild type to less than 1% of control levels. It also injured cell membranes, inhibited dehydrogenase activity, altered pigment composition, and reduced activity of photosystem II. Membrane damage was assessed through measuring electrical conductivity and potassium content, dehydrogenase activity by degree of conversion of 2,3,5-triphenyl tetrazolium chloride to red triphenylformazan, and functioning of PS II by chlorophyll a fluorescence. In respect to most parameters the tolerant strain was usually less affected by copper than the wild strain and, in some cases, not at all. We demonstrated by inductively coupled plasma atomic emission spectrometry and energy-dispersive X-ray microanalysis that enriched copper in the medium resulted in increased uptake by both wild and tolerant photobiont cells, but extracellular concentrations were significantly higher in the tolerant strain. This suggested that, at least in vitro, extracellular deposition was a mechanism of copper tolerance.

Adaptation, Physiological↗
PubMed2005

Late type of daughter cell wall synthesis in one of the Chlorellaceae, Parachlorella kessleri (Chlorophyta, Trebouxiophyceae).

Autosporulation is a common mode of propagation for unicellular algae. Autospore-forming species of Chlorellaceae, Chlorella vulgaris Beijerinck, C. sorokiniana Shihira et Krauss, C. lobophora Andreyeva, and Parachlorella kessleri (Fott et Nováková) Krienitz et al. have glucosamine as the main constituent of their rigid cell wall. Recent phylogenetic analyses have showed that the Chlorellaceae divided into two sister groups: the Chlorella-clade and the Parachlorella-clade. We compared the cell wall structure and synthesis of the daughter cell wall in the four species by electron microscopy using rapid freezing and freeze substitution methods. The cell wall of C. vulgaris, C. sorokiniana, and C. lobophora consisted of an electron-dense thin layer with an average thickness of 17-20, 22, and 19 nm, respectively. In these three species, daughter cell wall synthesis occurred on the outer surface of the plasma membrane in the early cell-growth phase. The cell wall of P. kessleri, however, was electron-transparent and 54-59 nm in thickness. Ruthenium red staining of P. kessleri indicated that ruthenium-red-specific polysaccharides accumulated over the outer surface of the plasma membrane. Immunoelectron microscopic observation with an anti-beta-1, 3-glucan antibody and staining with wheat germ agglutinin (WGA) indicated that the cell wall contained beta-1, 3-glucan and WGA specific N-acetyl-beta-D-glucosamine. In P. kessleri, daughter cell wall synthesis began after successive protoplast division. The daughter cell wall synthesis during autosporulation in the four species of Chlorellaceae can be classified into two types-the early and the late types.

Cell Cycle↗
PubMed2001

Non-photosynthetic enhancement of growth by high CO2 level in the nitrophilic seaweed Ulva rigida C. Agardh (Chlorophyta).

The effects of increased CO2 levels (10,000 microl l(-1)) in cultures of the green nitrophilic macroalga Ulva rigida C. Agardh were tested under conditions of N saturation and N limitation, using nitrate as the only N source. Enrichment with CO2 enhanced growth, while net photosynthesis, gross photosynthesis, dark respiration rates and soluble protein content decreased. The internal C pool remained constant at high CO2, while the assimilated C that was released to the external medium was less than half the values obtained under ambient CO2 levels. This higher retention of C provided the source for extra biomass production under N saturation. In N-sufficient thalli, nitrate-uptake rate and the activity of nitrate reductase (EC 1.6.6.1) increased under high CO2 levels. This did not affect the N content or the internal C:N balance, implying that the extra N-assimilation capacity led to the production of new biomass in proportion to C. Growth enhancement by increased level of CO2 was entirely dependent on the enhancement effect of CO2 on N-assimilation rates. The increase in nitrate reductase activity at high CO2 was not related to soluble carbohydrates or internal C. This indicates that the regulation of N assimilation by CO2 in U. rigida might involve a different pathway from that proposed for higher plants. The role of organic C release as an effective regulatory mechanism maintaining the internal C:N balance in response to different CO2 levels is discussed.

Algal Proteins↗
PubMed1999

Photoadaptation of two members of the Chlorophyta (Scenedesmus and Chlorella) in laboratory and outdoor cultures: changes in chlorophyll fluorescence quenching and the xanthophyll cycle.

The role of the xanthophyll cycle in the adaptation of two chlorococcal algae Scenedesmus quadricauda and Chlorella sorokiniana to high irradiance was studied under laboratory and outdoor conditions. We wished to elucidate whether the xanthophyll cycle plays a key role in dissipating the excesses of absorbed light, as in higher plants, and to characterise the relationship between chlorophyll fluorescence parameters and the content of xanthophyll-cycle pigments. The xanthophyll cycle was found to be operative in both species; however, its contribution to overall non-photochemical quenching (NPQ) could only be distinguished in Scenedesmus (15-20% of total NPQ). The Scenedesmus cultures showed a larger pool of xanthophyll-cycle pigments than Chlorella, and lower sensitivity to photoinhibition as judged from the reduction of maximum quantum yield of photosystem II. In general, both algae had a larger xanthophyll-cycle pool when grown outdoors than in laboratory cultures. Comparing the two species, Scenedesmus exhibited a higher capacity to adapt to high irradiance, due to an effective quenching mechanism and high photosynthetic capacity; in contrast, Chlorella represents a species with a larger antennae system, less-efficient quenching and lower photosynthetic performance. Non-photochemical quenching (NPQ) induced through the xanthophyll cycle can, to a limited extent, represent a regulatory factor in diluted algal cultures grown in outdoor solar photobioreactors, as well as in natural algal phytoplankton populations exposed transiently to high irradiance. However, it does not play an appreciable role in dense, well-mixed microalgal suspensions.

Adaptation, Physiological↗
PubMed1996

Cell-wall polysaccharides from the marine green alga Ulva "rigida" (Ulvales, Chlorophyta)--NMR analysis of ulvan oligosaccharides.

In order to obtain information on sugar sequences in the water soluble polysaccharides from Ulva "rigida", oligosaccharides were purified by anion exchange and gel permeation chromatography from the partial acid hydrolysate of the native or desulfated ulvan. The chemical structure and sugar sequence of these oligomers were determined by 1H and 13C NMR spectroscopy to be alpha-L-rhamnosyl (1-->4) D-xylose, beta-D-glucuronosyluronic acid (1-->2)-alpha-L-rhamnosyl (1-->4) D-xylose, beta-D-glucuronosyluronic acid (1-->4) L-rhamnose 3 sulfate, beta-D-glucuronosyluronic acid (1-->4) [beta-D-glucuronosyluronic acid (1-->2)] L-rhamnose and beta-D-glucuronosyluronic acid (1-->4) [beta-D-glucuronosyluronic acid (1-->2)] alpha-L-rhamnosyl (1-->4) D-xylose. The sugar linkages and particularly the branching and the sulfate position were in total agreement with previous chemical results.

Carbohydrate Conformation↗
PubMed1985

The concept of the genus Ulothrix (Chlorophyta) strengthened by comparative cytology.

The revelation of the ultrastructure of the pyrenoid, flagellar apparatus and mitosis/cytokinesis in the green algal genus Ulothrix, in its traditional broad sense, resulted in the assignment of species to three newly defined green algal classes. The largest group consisting of 11 species (including the type species Ulothrix tenuissima) is classified in the Ulvophyceae, the species U. verrucosa and U. crenulata are transferred to the genus Klebsormidium in the Charophyceae while Ulothrix species with pointed apical cells (e.g. U. belkae) appear to belong to the Chlorophyceae. It is shown that cytokinesis in Ulothrix sensu stricto may vary from a strict furrowing mechanism in U. mucosa to a pattern in which the centripetal ingrowth is combined with a cell plate-like structure in U. palusalsa and U. zonata. Based on the growth habit, the type of life history and the ultrastructural features pertaining to cytokinesis and the flagellar apparatus, phylogenetic trends are suggested for the Ulotrichales sensu lato.

Cell Division↗
PubMed2006

Physiological characterization of Dunaliella sp. (Chlorophyta, Volvocales) from Yucatan, Mexico.

Physiological responses of Dunaliella salina and Dunaliella viridis, isolated from solar saltworks on the Yucatan Peninsula, were studied. Optimal growth temperature for D. salina was 22 degrees C (3.06 x 10(6) cells mL(-1)) and 26 degrees C for D. viridis (4.04 x 10(6)cells mL(-1)). Total carotenoid content in D. salina increased with temperature to a maximum of 35.14 pg cell(-1) at 38 degrees C. Dunaliella salina alpha-carotene and beta-carotene content was 0.083+/-0.003 and 0.598+/-0.020 mg 100g dry wt(-1) respectively, whereas lower values were found in D. viridis cultured under same experimental conditions (0.018+/-0.002 and 0.136+/-0.012 mg 100g dry wt(-1) respectively). The highest specific growth rate in D. salina was obtained at 10% NaCl (0.28 d(-1)), while its cell volume increased from 524 to 2066.93 microm(3) when cultured from 10% to 35% NaCl. Maximum photosynthetic rates were attained when increasing from optimal growing temperature to 30 degrees C for D. viridis (108 n mol O(2)microg chl alpha h(-1)) and D. salina (139 n mol O(2)microg chl alpha h(-1)). Photosynthetic responses to temperature variations indicated physiological adjustments in both species, with higher acclimation in D. salina. Evaluation of physiological attributes of these species will be used for to carry out mass cultivation.

Analysis of Variance↗
PubMed2006

Structure of a highly pyruvylated galactan sulfate from the Pacific green alga Codium yezoense (Bryopsidales, Chlorophyta).

A polysaccharide fraction consisting of d-galactose, sulfate, and pyruvate in a molar proportion of 4:2:1 was isolated from the green seaweed Codium yezoense by water extraction followed by ion-exchange chromatography. To elucidate its structure, modified polysaccharides were prepared by desulfation, depyruvylation, and by total removal of non-carbohydrate substituents. Structures of the native polysaccharide and of the products of its chemical modifications were investigated by methylation analysis as well as by 1D and 2D (1)H and (13)C NMR spectroscopy. The polysaccharide devoid of sulfate and pyruvate was subjected to two subsequent Smith degradations to afford a rather low-molecular and essentially linear (1-->3)-beta-d-galactan. A highly ramified structure was suggested for the native polysaccharide, which contains linear backbone segments of 3-linked beta-d-galactopyranose residues connected by (1-->6) linkages, about 40% of 3-linked residues being additionally substituted at C-6, probably by short oligosaccharide residues also containing (1-->3) and (1-->6) linkages. Sulfate groups were found mainly at C-4 and in minor amounts at C-6. Pyruvate was found to form mainly five-membered cyclic ketals with O-3 and O-4 of the non-reducing terminal galactose residues. The minor part of pyruvate forms six-membered cyclic ketals with O-4 and O-6. The absolute configurations of ketals (R for six-membered ketals and S for five-membered ones) were established using NMR spectral data.

Carbohydrate Sequence↗
PubMed2005

Pathogenesis of Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) in susceptible noctuid larvae.

Helicosporidium sp. is a unique, achlorophyllous green alga that has been reported to infect various insect orders, including Lepidoptera, Diptera, and Coleoptera. The infectious cyst stage is ingested by the host, ruptures in the midgut lumen, and releases a filamentous cell. Histopathological examinations using larvae of a susceptible noctuid host, Spodoptera exigua, showed both cysts and filamentous cells affiliated with the microvillar lining of the midgut epithelium. A considerable proportion of the ingested cysts (22-39%) were recovered in feces collected 24 h after ingestion. A small number of filamentous cells passed the midgut epithelium and entered the hemocoel within 4-24 h after cyst ingestion. After 48 h, vegetative cell stages were detected in the hemolymph, followed by a 4- to 5-day period of increasing multiplication. Cyst differentiation in the colonized hemolymph began 6-7 days after the treatment.

Animals↗
PubMed2003

Photobiological characteristics and photosynthetic UV responses in two Ulva species (Chlorophyta) from southern Spain.

The effect of different wavebands of artificial UV (UVB and UVA) and photosynthetically active radiation (PAR) was assessed in two species of the genus Ulva, U. olivascens and U. rotundata, from southern Spain in order to test for possible differences in acclimation of photosynthesis. Both species share similar morphology but are subject to different light environments: U. rotundata is an estuarine alga, inhabiting subtidal locations, while U. olivascens is an intertidal, sun-adapted organism. Algae were exposed to three different UV conditions, PAR+UVA+UVB, PAR+UVA and PAR for 7 d. Short-term exposure (6 h) was also carried out, using two PAR levels, 150 and 700 micromolm(-2)s(-1). Pigment contents and photosynthesis vs. irradiance curves from oxygen evolution were used to contrast sun- and shade adaptation between these species. O2-based net photosynthesis (Pmax) and PAM-chlorophyll fluorescence (optimal quantum yield, Fv/Fm) were used as parameters to evaluate photoinhibition of photosynthesis in the experiments. The results underline different photobiological characteristics among species: the subtidal U. rotundata had higher contents of pigments (Chl a, Chl b and carotenoids) than the sun-adapted U. olivascens, which resulted in higher thallus absorptance and P-I parameters characterized by higher photosynthetic efficiency at limiting irradiances (alpha) and lower saturating points for photosynthesis (Ek). After 7 d exposure, photoinhibition of Fv/Fm was close to 40-45% in both species. Differences between UV treatments were seen in U. rotundata after 5 d and after 7 d in U. olivascens, in which PAR+UVA impaired strongly photosynthesis (80%). Such patterns were correlated with a progressive decrease in pigment contents, specially chlorophylls. In short-term (6 h) exposures, combinations of UVA+UVB and high PAR level resulted in high rates of photoinhibition of chlorophyll fluorescence (68-92%) in U. rotundata, whereas in U. olivascens photoinhibition ranged between 42% and 53%. Photoinhibition under low PAR combined to UV radiation was lower than observed under high PAR. Net O2-Pmax revealed similar response among the species, with maximal photoinhibition rates close to 60% in algae incubated under high PAR+UVA+UVB. In the case of UV exposure in combination with low PAR, the highest photoinhibition rates were measured in U. rotundata.

Carotenoids↗
PubMed2006

PCR-identification of Dunaliella salina (Volvocales, Chlorophyta) and its growth characteristics.

The saline pond microalga, Dunaliella salina (Dunal) Teod. maintained in De Walne's (basal) medium under laboratory conditions was confirmed by amplifying the chromosomal DNA of the microalga by PCR with specific primers MA1 and MA2. Seaweed extracts obtained from Sargassum wightii and Ulva lactuca were amended separately at 1.0%, 1.5%, 2.0% and 2.5% levels to the basal medium in order to assess their potential on the growth and concentration of pigments, viz. Chl a, Chl b and beta-carotene of the alga. beta-Carotene was isolated and visible absorption spectrum was taken at 443 and 475 nm confirmed the presence of 9-cis-beta-carotene and all-trans-beta-carotene isomers. Maximum yield, highest division rate (mu) and highest pigment concentrations were observed in the cells grown in 1.5% S. wightii and 2.0% U. lactuca amended medium and these cells were subjected to DAPI staining. The results of epifluorescence microscopy and image analysis revealed a significant enhancement of the cell and nuclear area of the microalgae.

Chlorophyll↗
PubMed2006

Protective effect of Dunaliella salina (Volvocales, Chlorophyta) against experimentally induced fibrosarcoma on wistar rats.

The beta-carotene-yielding microalga, Dunaliella salina (Dunal) Teod. maintained in De Walne's medium was harvested and lyophilized. Fibrosarcoma was induced in rats by 20-methylcholanthrene. 0.5 g and 1.0 g of lyophilized D. salina powder was administered to the rats orally through carboxy methyl cellulose. Cisplatin was administered along with vitamin E to compare the protective effect of D. salina against fibrosarcoma. Administration of D. salina decreased the levels of cholesterol and lactate dehydrogenase as well as the activities of catalase, superoxide dismutase, serum aspartate aminotransaminase, serum alanine aminotransferase, when compared to control. A significant reduction in the levels of hepatic and renal RNA and DNA was observed in the sarcoma rats when treated with D. salina powder. Histopathological studies of tumor tissues showed regenerative and regressive changes. beta-carotene globules isolated from the powder of Dunaliella salina confirmed the presence of 9-cis-beta-carotene and all-trans-beta-carotene.

Alanine Transaminase↗